Lipofectamine

Lipofectamine is a transfection reagent used to introduce exogenous DNA or RNA into cells through lipofection. This is a chemical method that relies on the formation of a liposome, from Lipofectamine lipid subunits, around a transfection payload, which through fusion with the cellular membrane, then enters target cells. This fusion process is facilitated by the positively charged surface of the liposome, which interacts with negatively charged cell membranes. (Cardarelli et al., 2016; Hasan et al., 2021)

Chemical transfection methods, such as Lipofectamine, have several pros and cons when compared to physical methods, such as electroporation. For instance, one advantage of Lipofectamine is that it causes minimal cell damage. This is because delivery of nucleic material occurs through fusion with cell membranes, rather than forced permeabilization through high voltage electric pulses, as in electroporation. On the other hand, one disadvantage of Lipofectamine, and many chemical transfection methods in general, is that the transfection efficiency is lower. (Sharifi et al., 2015; Yamano et al., 2010)

Lipofectamine is available in multiple different formats, including Lipofectamine 2000, Lipofectamine 3000, Lipofectamine RNAiMAX and Lipofectamine LTX. While it is difficult to state the best Lipofectamine, since this highly depends on the experimental requirements, there are some general considerations. If transfection efficiency is of concern, Lipofectamine 3000 is better than Lipofectamine 2000, with average efficiency roughly 70% vs 50% respectively, depending on cell type. Pricing-wise, however, Lipofectamine 3000 is more costly per unit than the 2000 variant. For siRNA and miRNA delivery into cells, Lipofectamine RNAiMAX is recommended. Finally, Lipofectamine LTX is recommended for plasmid transfection. (Daneshvar et al., 2014)

While the specific usage of Lipofectamine should be optimized for particular experimental conditions, there are some general guidelines to follow. For instance, when using Lipofectamine 300, ~100 ng of DNA should be added per well in a 96-well plate. Incubation with the reagent should be 10 to 15 minutes at room temperature. After addition of transfection payload, cells should be incubated 2 to 4 days at 37 °C before analysis. Finally, for storage, the Lipofectamine 3000 reagent can be stored for 12 months at 2 to 8 °C.

Cardarelli, F., Digiacomo, L., Marchini, C., Amici, A., Salomone, F., Fiume, G., ... & Caracciolo, G. (2016). The intracellular trafficking mechanism of Lipofectamine-based transfection reagents and its implication for gene delivery. Scientific reports, 6(1), 25879.

Daneshvar, N., Rasedee, A., Mehrbod, P., & Hashem Boroojerdi, M. (2014). Optimizing transfection of umbilical cord mesenchymal stem cells utilizing minicircle plasmid/lipofectamine LTX complex. Journal of Applied Biotechnology Reports, 1(3), 111-116.

Hasan, M. M., Ragnarsson, L., Cardoso, F. C., & Lewis, R. J. (2021). Transfection methods for high-throughput cellular assays of voltage-gated calcium and sodium channels involved in pain. Plos one, 16(3), e0243645.

Sharifi Tabar, M., Hesaraki, M., Esfandiari, F., Sahraneshin Samani, F., Vakilian, H., & Baharvand, H. (2015). Evaluating electroporation and lipofectamine approaches for transient and stable transgene expressions in human fibroblasts and embryonic stem cells. Cell Journal (Yakhteh), 17(3), 438-450.

Yamano, S., Dai, J., & Moursi, A. M. (2010). Comparison of transfection efficiency of nonviral gene transfer reagents. Molecular biotechnology, 46, 287-300.